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首页> 外文期刊>Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society >Stromal cell-derived factor 1-alpha (SDF)-induced human T cell chemotaxis becomes phosphoinositide 3-kinase (PI3K)-independent: role of PKC-theta.
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Stromal cell-derived factor 1-alpha (SDF)-induced human T cell chemotaxis becomes phosphoinositide 3-kinase (PI3K)-independent: role of PKC-theta.

机译:基质细胞衍生的因子1-alpha(SDF)诱导的人类T细胞趋化性成为磷酸肌醇3激酶(PI3K)独立的:PKC-θ的作用。

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Stromal cell-derived factor 1alpha (SDF-1alpha) is the exclusive ligand for the chemokine receptor CXCR4. This receptor plays a pivotal role in immune responses, the pathogenesis of infection such as HIV, and cellular trafficking. However, the signaling mechanisms regulating SDF-driven T cell migration are not well defined. In this study, we determined the role of PI3K and protein kinase C- theta (PKC-theta) in SDF-induced human T cell migration in fresh versus cultured T cells. Purified human T cells (fresh vs. 48 h in media, unstimulated or activated by anti-CD3+anti-CD28) were used. Western blots showed that SDF induced phospho-(p)-Akt [threonine (Thr)308 and serine 473], a proxy for PI3K activity, in fresh cells and p-PKC-theta in 48 h unstimulated cells. LY294002 (PI3K inhibitor) reduced SDF-induced chemotaxis in fresh cells by 51%, whereas it minimally affected chemotaxis in 48 h unstimulated or activated cells. However, a specific PKC-theta inhibitor, pseudosubstrate for PKC-theta, reduced chemotaxis in 48 h unstimulated and stimulated T cells by 72% and 87%, respectively. Thus, chemotaxis becomes independent of PI3K signaling in human T cells cultured for 48 h. Under these conditions, PKC-theta is phosphorylated (Thr538) by SDF, and chemotaxis becomes largely PKC-theta-dependent.
机译:基质细胞衍生因子1alpha(SDF-1alpha)是趋化因子受体CXCR4的唯一配体。该受体在免疫应答,HIV等感染的发病机理和细胞运输中起关键作用。但是,调节SDF驱动的T细胞迁移的信号传导机制尚不明确。在这项研究中,我们确定了PI3K和蛋白激酶C-theta(PKC-theta)在SDF诱导的人T细胞在新鲜T细胞与培养T细胞中的迁移中的作用。使用纯化的人T细胞(在培养基中相对于48小时新鲜,未受抗CD3 +抗CD28刺激或激活)。 Western印迹显示,SDF诱导了新鲜细胞和48小时未刺激细胞中p-PKC-theta的磷酸化-(p)-Akt [苏氨酸(Thr)308和丝氨酸473],这是PI3K活性的代表。 LY294002(PI3K抑制剂)将SDF诱导的新鲜细胞趋化性降低51%,而在48小时未刺激或活化的细胞中,其对趋化性的影响最小。但是,一种特定的PKC-theta抑制剂,即PKC-theta的假底物,在48小时内未刺激和刺激的T细胞的趋化性分别降低了72%和87%。因此,趋化性变得独立于培养48小时的人类T细胞中的PI3K信号传导。在这些条件下,PKC-θ被SDF磷酸化(Thr538),趋化性很大程度上取决于PKC-θ。

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